A diarylheptanoid compound from Alpinia officinarum Hance ameliorates high glucose-induced insulin resistance by regulating PI3K/AKT-Nrf2-GSK3β signaling pathways in HepG2 cells
A diarylheptanoid compound from Alpinia officinarum Hance ameliorates high glucose-induced insulin resistance by regulating PI3K/AKT-Nrf2-GSK3β signaling pathways in HepG2 cells
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来自高良姜 Hance 的二芳基庚烷化合物通过调节 HepG2 细胞中的 PI3K/AKT-Nrf2-GSK3β 信号通路改善高葡萄糖诱导的胰岛素抵抗
DOI：
10.1016/j.jep.2022.115397
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发表时间：
2022
影响因子：
5.4
通讯作者：
Jun-qing Zhang
中科院分区：
文献类型：
--
作者：
Xu-guang Zhang;Ai-xia Liu;Yu-xin Zhang;Ming-yan Zhou;Xiang-yi Li;Ming-hai Fu;Yi-peng Pan;Jian Xu;Jun-qing Zhang
高良姜（Alpinia officinarum Hance）是一种多年生天然药食两用草本植物，数千年来一直被传统地用于治疗感冒、胃痛和糖尿病。1，7-二苯基-4E-烯-3-庚酮（DPH_5）是一种从黄芪（A.为探讨DPH 5对胰岛素抵抗（IR）的保护作用及其可能的作用机制，以HepG 2细胞为研究对象，研究DPH 5对IR的保护作用。采用流式细胞术检测高糖诱导的胰岛素抵抗HepG 2细胞的葡萄糖摄取和活性氧（ROS）水平。采用相应的检测试剂盒分析葡萄糖消耗量以及丙二醛（MDA）和超氧化物歧化酶（SOD）水平。与胰岛素信号传导、葡萄糖代谢和抗氧化因子相关的mRNA和蛋白质的表达，包括胰岛素受体底物-1（IRS 1）、磷脂酰肌醇3-激酶（PI 3 K）、蛋白激酶B（AKT）、葡萄糖转运蛋白-4、糖原合成酶激酶-3 β（GSK 3 β）、葡萄糖激酶（GCK）、丙酮酸激酶（PK）、磷酸烯醇式丙酮酸羧激酶（PEPCK）、采用实时定量聚合酶链反应和蛋白质印迹法测定葡萄糖-6-磷酸酶（G6 β）、核因子-红细胞2相关因子2（Nrf 2）、血红素加氧酶-1（HO-1）、NADPH醌氧化还原酶（NQO 1）和谷胱甘肽过氧化物酶（GSH-Px）。进一步通过分子对接分析DPH 5对PI 3 K、AKT、Nrf 2和GSK 3 β等关键靶点的作用机制，发现DPH 5可改善胰岛素抵抗细胞的胰岛素抵抗，主要表现为葡萄糖摄取和葡萄糖消耗增加。此外，DPH 5还可通过激活Nrf 2/HO-1元件增强抗氧化能力，包括增加Nrf 2、HO-1、SOD、NQO 1和GSH-Px的表达，降低MDA、ROS和JNK水平，从而改善氧化应激，最终缓解IR。此外，DPH 5还可促进IRS 1、PI 3 K、AKT、GSK 3 β、GCK和PK的表达，下调PEPCK和G6 pase的表达。从而加速葡萄糖利用并增强胰岛素敏感性。DPH 5减轻IR的作用机制与PI 3 K/AKT和Nrf 2/HO-1介导的GSK 3 β信号通路调节有关，特异性抑制剂LY 294002和ML 385进一步证实了这一结果。分子对接结果表明，DPH 5与PI 3 K、AKT、Nrf 2和GSK 3 β之间存在不同的调控位点和相互作用力; DPH 5通过调节PI 3 K/AKT-Nrf 2-GSK 3 β通路改善氧化应激和葡萄糖代谢，从而改善IR。我们的研究结果表明DPH 5作为治疗2型糖尿病的天然药物的潜力。
Alpinia officinarum Hance, a perennial natural medicine-food herb, has been traditionally used to treat colds, stomachache, and diabetes for thousands of years. 1,7-Diphenyl-4E-en-3-heptanone (DPH5), a diarylheptanoid isolated from the rhizome of A. officinarum has been reported to be safe and to have antioxidant and hypoglycemic effects, suggesting its potential in the treatment of insulin resistance (IR).Aim of to investigate the protective effect of DPH5 on IR and elucidate its underlying mechanism of action.HepG2 cells were used as the research objects. Glucose uptake and reactive oxygen species (ROS) levels in high glucose-induced insulin-resistant HepG2 cells were assessed using flow cytometry. Glucose consumption and the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were analyzed using the corresponding assay kits. The expression of mRNA and proteins related to insulin signaling, glucose metabolism, and antioxidant factor, including insulin receptor substrate-1 (IRS1), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), translocation of glucose transporter-4, glycogen synthase kinase-3β (GSK3β), glucokinase (GCK), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), nuclear factor-erythroid 2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH quinoneoxidoreductase (NQO1), and glutathione peroxidase (GSH-Px) was determined using real-time quantitative polymerase chain reaction and western blotting. Furthermore, molecular docking was performed to determine the spatial mechanism of DPH5 on the key targets PI3K, AKT, Nrf2, and GSK3β.DPH5 could improve IR that manifested as increased glucose uptake and glucose consumption in insulin-resistant HepG2 cells. Moreover, DPH5 could enhance antioxidant capacity by activating Nrf2/HO-1 elements, including increasing Nrf2, HO-1, SOD, NQO1, and GSH-Px expression and reducing MDA, ROS, and JNK levels, thereby improving oxidative stress and ultimately alleviating IR. Additionally, DPH5 could promote the expression of IRS1, PI3K, AKT, GSK3β, GCK, and PK, and downregulate the expression of PEPCK and G6pase, thereby accelerating glucose utilization and enhancing insulin sensitivity. The mechanism underlying the effect of DPH5 in alleviating IR was related to the PI3K/AKT- and Nrf2/HO-1-mediated regulation of the GSK3β signaling pathway, and the results were further confirmed using the specific inhibitors LY294002 and ML385. Results from molecular docking indicated that there were different regulatory sites and interacting forces between DPH5 and PI3K, AKT, Nrf2, and GSK3β; however, the binding force was relatively strong.DPH5 improved oxidative stress and glucose metabolism via modulating the PI3K/AKT-Nrf2-GSK3β pathway, thereby ameliorating IR. Overall, our findings suggest the potential of DPH5 as a natural medicine to treat type-2 diabetes mellitus.